vector with tag recommendation - (Nov/23/2005 )

I want to make a clone with some tag, so does anyone recommend any vector for this purpose, and where to buy this vector? as i know, HA-tag and Myc-tag vector are available in clontech, how about other tags? which one is better? thanks.

--Jason

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FLAG is an awesome tag (n-terminus, seven residues, high affinity), but it's expensive (for antibodies). His tag antibodies are cheaper but the binding affinity is not as good.

There are so many vectors with binding affinity tags that you are best off finding the one that is easiest to acquire (i.e. FREE) and use. pET (Novagen) and Gateway vectors (Invitrogen) both have their own set of tags.

-Matt

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FLAG is an awesome tag (n-terminus, seven residues, high affinity), but it's expensive (for antibodies). His tag antibodies are cheaper but the binding affinity is not as good.

There are so many vectors with binding affinity tags that you are best off finding the one that is easiest to acquire (i.e. FREE) and use. pET (Novagen) and Gateway vectors (Invitrogen) both have their own set of tags. If you are smart, you can create your own tags in your own vectors through an addition by inverse PCR.

-Matt

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hi
i like pFHM vectors which contains Flag myc and Ha tags.

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thanks, so which company has this powerful vector??

--Jason

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well
i have it in my lab. but i don't know if it's an home made or not. that's wh i didn't mentioned the comany

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I have checked that Sigma has a mammanlian expression vector with both 3xFLAG and c-Myc tags, but as HA tag should be a patent for clontech only, so urs may be a home-made one, just guess.

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well i asked and you guessed good. We started from a flagplasmid and cloned into it ha and myc. 3flag myc vectors seems very good actually, doesn't it?
i can send you the sequence i cloned if you want

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hi jasonwu,

what i've done in my lab is to engineer flag/his tag in the c-terminal end and use invitrogen's gateway system to transfer the gene into appropriate vector. for example, transer GOI-flag/his clone into pDEST15 which contains GST tag on the N-terminal end for bacterial expression therefore, you have GST-GOI-Flag/his. if your gene-of-interest causes toxicity to the bacterial cells then simply transfer it into pDEST20 (i believe) for baculo vector.

i've been using gateway system for about 4 yrs now and never looked back. system is great because of it's simplicity and flexiblity. try it!!

good luck,

steven

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Hi!!!

Steven, did you include some buffer amino acids between flag/his and your gene of interest??? I just directly fused flag to my gene of interest and didn't transfer it into expression vector yet. I’m wondering whether it is going to work or not.

Greetings,
karmen

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